Proteomic identification of oxidative-stress-reporting biomarkers differentially secreted from human neuroblastoma SH-SY5Y cells

Abstract

The free-radical theory predicts that the oxidative stress accelerates the rate of aging and increases the onset of degenerative disorders in the elderly. Dopaminergic neurons are especially vulnerable to age-related neuronal disorders due to reactive oxygen species generated in the pathway of dopamine metabolism. Biochemical changes occurring in substantia nigra of Parkinson's disease patients suggest that the oxidative-stress-induced cell damages may be involved in the neurodegeneration. In our previous researches, we found that the dephosphorylation of elongation factor-2 and phosphorylation of nuclear lamin A/ C might be neuronal cell specific response to oxidative stress. (Nakamura et al. BBA, 1763(9), 977-989, 2006) The dephosphorylation and phosphorylation of those proteins are significant biomarkers for analyzing the molecular mechanisms of the stress response, however, such a phosphoproteome analysis is thought to be inappropriate for clinical investigation of neurodegeneration if it was not detectable in cerebrospinal fluid or serum of patients. Thus, we proceeded to the 2D-DIGE analysis of secretome, proteome of secreted proteins, using the culture system in which oxidative stress was applied to human SH-SY5Y neuroblastoma cells. As the result of our secretome analysis, we identified ubiquitin-activating enzyme E1, ubiquitin-conjugating enzyme E2 N, ubiquitin C-terminal hydrorase-L1, 14-3-3 protein isoforms, Rab GDP dissociation inhibitor β, Rho GDP-dissociation inhibitor 1, peroxiredoxin-2, glutathione S-transferase P, α enolase, LDH B chain as oxidative-stress-reporting biomarker candidates.

Effects of Mercury onDictyostelium discoideum: Proteomics Reveals the Molecular Mechanisms of Physiological Adaptation and Toxicity

“…In particular GDI retrieves GDP-bearing rab proteins from target membranes after a vesicular transport event. An increment of secreted rab GDP dissociation inhibitor has been recently reported as a biomarker of oxidative stress by 2-DDIGE in human neuroblastoma SH-SY5Y cells …”

“…An increment of secreted rab GDP dissociation inhibitor has been recently reported as a biomarker of oxidative stress by 2-DDIGE in human neuroblastoma SH-SY5Y cells. 99 In 10 µM Hg exposed amoebae there is a strong increase of ROS; it appears of interest to clarify to what extent the production of oxyradicals has affected the proteome of the soluble proteins. We evaluated spectrophotometrically the level of protein carbonylation in 2 µM and 10 µM treated amoebae and results reveal a significant increase in carbonylation only after the treatment with 10 µM mercury.…”

Functions of Tat: the versatile protein of human immunodeficiency virus type 1

“…Oxidative stress induced by ROS is suspected to be a major cause of chronic neurodegenerative diseases (Toda et al, 2007). Oxidative stress has been demonstrated both in the brain and in cerebrospinal fluid (CSF) of HAD patients (Turchan et al, 2003).…”